Fluorimetric estimation of the concentration of NADH from human blood samples

We report the quantitative fluorimetric estimation of the concentration of NADH in human blood samples. To measure both free and bound NADH, the 340-nm-excited fluorescence spectra was recorded from blood samples to which SDS was added. Diffuse reflectance and the transmittance spectra of the same sample were also recorded over the wavelength range 340-650 nm. These measurements were used to correct the measured fluorescence spectra for the wavelength-dependent reduction in fluorescence due to absorption and scattering in the blood sample. NADH concentrations estimated using the corrected fluorescence spectra were found to be in reasonable agreement with the concentrations estimated by the biochemical method.